Membrane-Pore Forming Characteristics of the Bordetella pertussis CyaA-Hemolysin Domain
AbstractPreviously, the 126-kDa Bordetella pertussis CyaA pore-forming/hemolysin (CyaA-Hly) domain was shown to retain its hemolytic activity causing lysis of susceptible erythrocytes. Here, we have succeeded in producing, at large quantity and high purity, the His-tagged CyaA-Hly domain over-expressed in Escherichia coli as a soluble hemolytically-active form. Quantitative assays of hemolysis against sheep erythrocytes revealed that the purified CyaA-Hly domain could function cooperatively by forming an oligomeric pore in the target cell membrane with a Hill coefficient of ~3. When the CyaA-Hly toxin was incorporated into planar lipid bilayers (PLBs) under symmetrical conditions at 1.0 M KCl, 10 mM HEPES buffer (pH 7.4), it produced a clearly resolved single channel with a maximum conductance of ~35 pS. PLB results also revealed that the CyaA-Hly induced channel was unidirectional and opened more frequently at higher negative membrane potentials. Altogether, our results first provide more insights into pore-forming characteristics of the CyaA-Hly domain as being the major pore-forming determinant of which the ability to induce such ion channels in receptor-free membranes could account for its cooperative hemolytic action on the target erythrocytes. View Full-Text
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Kurehong, C.; Kanchanawarin, C.; Powthongchin, B.; Katzenmeier, G.; Angsuthanasombat, C. Membrane-Pore Forming Characteristics of the Bordetella pertussis CyaA-Hemolysin Domain. Toxins 2015, 7, 1486-1496.
Kurehong C, Kanchanawarin C, Powthongchin B, Katzenmeier G, Angsuthanasombat C. Membrane-Pore Forming Characteristics of the Bordetella pertussis CyaA-Hemolysin Domain. Toxins. 2015; 7(5):1486-1496.Chicago/Turabian Style
Kurehong, Chattip; Kanchanawarin, Chalermpol; Powthongchin, Busaba; Katzenmeier, Gerd; Angsuthanasombat, Chanan. 2015. "Membrane-Pore Forming Characteristics of the Bordetella pertussis CyaA-Hemolysin Domain." Toxins 7, no. 5: 1486-1496.